Tangle formation and downregulation of miR‐132‐3p in LOAD A‐G Tangle formation and downregulation of miR‐132‐3p in LOAD A‐G.Downregulation of miR‐132‐3p colocalizes with hyper‐phosphorylated Tau. Tangle formation and downregulation of miR‐132‐3p in LOAD A‐G.Downregulation of miR‐132‐3p colocalizes with hyper‐phosphorylated Tau. (A) In situ hybridization of miR‐132‐3p (red) and immunohistochemical detection of hyper‐phosphorylated Tau (green, AT8 antibody) revealed that hyper‐phosphorylated Tau builds up in some miR‐132‐3p positive neurons of a BRV patient. Panels B, D, F depict tangle formation in layer III neurons (B: merge, D: AT8, F: miR‐132‐3p). Panels C, E, G depict tangle formation in layer V neurons (C: merge, E: AT8, G: miR‐132‐3p). Arrows denote examples of miR‐132‐3p and AT8 colocalization. Scale bar in A corresponds to 100 µm. Scale bar in G represents 50 µm.H.miR‐132‐3p expression is decreased in tangle‐bearing neurons. The average fluorescence levels of miR‐132‐3p in AT8(+) and AT8(−) neurons in layers III and V were quantified for one Braak V individual and two Braak VI subjects. In layer III, the average expression levels are based on 52, 81 and 190 AT8(−) neurons and 18, 20 and 35 AT8(+) neurons, respectively. In layer V, the average expression levels are based on 32, 45 and 145 AT8(−) neurons and 14, 17 and 41 AT8(+) neurons, respectively. Note the significant decrease in miR‐132‐3p levels in AT8(+) neurons in both cortical layers (p = 0.0003 for layer III and p = 0.0165 for layer V, paired Student's t‐test).I.Quantification of miR‐132‐3p expression levels in layers III and V neurons. The graph represents the miR‐132‐3p fluorescence intensity of all neurons found in panel B (layer III) and panel C (layer V). Note that miR‐132‐3p expression is significantly lower in layer V neurons of this BRV subject (p < 0.0001, Mann–Whitney U test). Red lines indicate median expression levels. Pierre Lau et al. EMBO Mol Med. 2013;5:1613-1634 © as stated in the article, figure or figure legend